Cryptic phosphoribosylase activity of NAMPT restricts the virion incorporation of viral proteins

Copyright and License

© 2024, The Author(s), under exclusive licence to Springer Nature Limited

Acknowledgement

We thank S.-I. Imai (Washington University) for providing the Nampt^fl/fl mice, S. Petteri (Stanford University) and N. Graham (University of Southern California) for LC–MS techniques, J. Hao (Poochon Scientific) for APEX-related protein identification, W. Beatty (Washington University) for electron microscopy analysis, W. Yuan (University of Southern California), D. Knipe (Harvard Medical School) and R. Longnecker (Northwestern University) for providing HSV-1 plasmids, J. Zhao (Cleveland Clinic Foundation) for guidance on generating HSV-1 mutant viruses, G. Cohen (University of Pennsylvania) for anti-gH antibody, C. Zhen (University of Calgary) for anti-VP22 antibody, B. Lomenick (CalTech) for processing MS samples, and N. Graham (University of Southern California) and S. Pitteri (Stanford University) for assistance on mass spectrometry analysis. We are grateful to Y. Zhou, S. Rice, J. Carriere and J. Xiao for their assistance. This work was partly supported by a startup fund from the Herman Ostrow School of Dentistry of the University of Southern California and grants from the National Institutes of Health (AG070904, CA285192 and AI180537) and Infectious Disease Society of America Foundation (Microbial Pathogenesis in AD) (P.F.) and National Natural Science Foundation of China (81821002) (C.H.).

Data Availability

All raw datasets used for phosphoribosylated peptide analysis can be found in the PRIDE public repository under project accession number PXD050684. All source data supporting the main figures and extended data figures are published within the paper. Source data are provided with this paper.

Supplemental Material

Supplementary Information
Supplementary Figures

Supplementary Table 1
Supplementary Table 1. An Excel file summarizing NAMPT-binding proteins identified by APEX2-mediated biotinylation and mass spectrometry, related to Fig. 3g (source file). Supplementary Table 2. An Excel file summarizing the NAMPT-binding HSV-1 proteins identified by APEX2-mediated biotinylation and mass spectrometry, related to Fig. 3g. Supplementary Table 3. An Excel file summarizing the phosphoribosylated peptides identified from a regular (2 h) run of HSV-1-infected cell lysates of HeLa expressing wildtype NAMPT or NAMPT-H247E mutant, related to Extended Data Fig. 6c and 6d. Supplementary Table 4. An Excel file showing the percentage of phosphoribosylated peptides normalized to the total peptides analyzed in Supplementary Table 3, related to Extended Data Fig. 6c and 6d. Supplementary Table 5. An Excel file providing the raw data for total peptides and proteins identified from an extended (4 h) run of HSV-1-infected cell lysates of HeLa expressing wildtype NAMPT or NAMPT-H247E mutant, related to Extended Data Fig. 6c and 6d. Supplementary Table 6. An Excel file providing the raw data for viral peptides and proteins identified from an extended (4 h) run of HSV-1-infected cell lysates of HeLa expressing wildtype NAMPT or NAMPT-H247E mutant, related to Extended Data Fig. 6c and 6d. Supplementary Table 7. An Excel file showing the percentage of phosphoribosylated viral peptides summarized in Supplementary Tables 5 and 6, related to Extended Data Fig. 6c and 6d. Supplementary Table 8. An Excel file summarizing the quantification of viral and cellular phosphoribosylated peptides summarized in Supplementary Tables 5 and 6, related to Extended Data Fig. 6c and 6d. Supplementary Table 9. An Excel file showing the percentage of phosphoribosylated total (viral and cellular) peptides summarized in Supplementary Tables 5 and 6, related to Extended Data Fig. 6c and 6d. Supplementary Table 10. A list of primers used in this study.